The World's #1 Renewable Energy Network for News, Information, and Companies.

Engineering Cells for More Efficient Biofuel Production

In the search for renewable alternatives to gasoline, heavy alcohols such as isobutanol are promising candidates. Not only do they contain more energy than ethanol, but they are also more compatible with existing gasoline-based infrastructure. For isobutanol to become practical, however, scientists need a way to reliably produce huge quantities of it from renewable sources.

MIT chemical engineers and biologists have now devised a way to dramatically boost isobutanol production in yeast, which naturally make it in small amounts. They engineered yeast so that isobutanol synthesis takes place entirely within mitochondria, cell structures that generate energy and also host many biosynthetic pathways. Using this approach, they were able to boost isobutanol production by about 260 percent.

Though still short of the scale needed for industrial production, the advance suggests that this is a promising approach to engineering not only isobutanol but other useful chemicals as well, says Gregory Stephanopoulos, an MIT professor of chemical engineering and one of the senior authors of a paper describing the work in the Feb. 17 online edition of Nature Biotechnology.

“It’s not specific to isobutanol,” Stephanopoulos says. “It’s opening up the opportunity to make a lot of biochemicals inside an organelle that may be much better suited for this purpose compared to the cytosol of the yeast cells.”

Stephanopoulos collaborated with Gerald Fink, an MIT professor of biology and member of the Whitehead Institute, on this research. The lead author of the paper is Jose Avalos, a postdoc at the Whitehead Institute and MIT.

Historically, researchers have tried to decrease isobutanol production in yeast, because it can ruin the flavor of wine and beer. However, “now there’s been a push to try to make it for fuel and other chemical purposes,” says Avalos, the paper’s lead author.

Yeast typically produce isobutanol in a series of reactions that take place in two different cell locations. The synthesis begins with pyruvate, a plentiful molecule generated by the breakdown of sugars such as glucose. Pyruvate is transported into the mitochondria, where it can enter many different metabolic pathways, including one that results in production of valine, an amino acid. Alpha-ketoisovalerate (alpha-KIV), a precursor in the valine and isobutanol biosynthetic pathways, is made in the mitochondria in the first phase of isobutanol production. 

Valine and alpha-KIV can be transported out to the cytoplasm, where they are converted by a set of enzymes into isobutanol. Other researchers have tried to express all the enzymes needed for isobutanol biosynthesis in the cytoplasm. However, it’s difficult to get some of those enzymes to function in the cytoplasm as well as they do in the mitochondria.  

The MIT researchers took the opposite approach: They moved the second phase, which naturally occurs in the cytoplasm, into the mitochondria. They achieved this by engineering the metabolic pathway’s enzymes to express a tag normally found on a mitochondrial protein, directing the cell to send them into the mitochondria. 

This enzyme relocation boosted the production of isobutanol by 260 percent, and yields of two related alcohols, isopentanol and 2-methyl-1-butanol, went up even more — 370 and 500 percent, respectively.

There are likely several explanations for the dramatic increase, the researchers say. One strong possibility, though difficult to prove experimentally, is that clustering the enzymes together makes it more likely that the reactions will occur, Avalos says.

Another possible explanation is that moving the second half of the pathway into the mitochondria makes it easier for the enzymes to snatch up the limited supply of precursors before they can enter another metabolic pathway.

“Enzymes from the second phase, which are naturally out here in the cytoplasm, have to wait to see what comes out of the mitochondria and try to transform that. But when you bring them into the mitochondria, they’re better at competing with the pathways in there,” Avalos says.

The findings could have many applications in metabolic engineering. There are many situations where it could be advantageous to confine all of the steps of a reaction in a small space, which may not only boost efficiency but also prevent harmful intermediates from drifting away and damaging the cell. 

The researchers are now trying to further boost isobutanol yields and reduce production of ethanol, which is still the major product of sugar breakdown in yeast. 

“Knocking out the ethanol pathway is an important step in making this yeast suitable for production of isobutanol,” Stephanopoulos says. “Then we need to introduce isobutanol synthesis, replacing one with the other, to maintain everything balanced within the cell.”

The research was funded by the National Institutes of Health and Shell Global Solutions.

Lead image: Lab flasks via Shutterstock

RELATED ARTICLES

UK Parliament Clean Energy Leaders

UK Government Names Clean Energy Cabinet Members

David Appleyard, Contributing Editor With the UK general election now over and a majority Conservative Party government in place, the re-elected Prime Minister David Cameron has now named key members of the government charged with steering the UK’s clean energ...
Microgrids

Coast to Coast and Across the Electric System, Microgrids Provide Benefits to All

Dick Munson, Environmental Defense Fund At the most obvious level, microgrids could disrupt today’s utilities and their regulated-monopoly business model, because they challenge the centralized paradigm. In a nutshell, microgrids are localized power grids that ha...
Alaska Airlines Biofuel

Alaska Airlines, Gevo To Demonstrate Renewable Alcohol-to-Jet Fuel

Jim Lane, Biofuels Digest

In Colorado, Gevo and Alaska Airlines announced a strategic alliance to purchase Gevo’s renewable jet fuel and fly the first-ever commercial flight on alcohol-to-jet fuel (ATJ).

Germany wind turbines. Credit: Shutterstock.

Germany's Powerhouse Feels Pinch of Merkel’s Shift to Renewables

Tino Andresen, Bloomberg North Rhine-Westphalia, the German state that’s home to utilities RWE AG and EON SE, is losing its standing as the country’s powerhouse as wind and solar energy begin to displace conventional sources. Electricity consumers ...

CURRENT MAGAZINE ISSUE

03/01/2015
Volume 18, Issue 3
file

STAY CONNECTED

To register for our free
e-Newsletters, subscribe today:

SOCIAL ACTIVITY

Tweet the Editors! @megcichon @jennrunyon

FEATURED PARTNERS



EVENTS

International Fuel Ethanol Workshop & Expo

Now in its 31st year, the FEW provides the global ethanol industry with...

Green Energy Summer School: Introduction to Bioenergy

This introductory course gives a detailed overview of the principles of ...

23rd European Biomass Conference and Exhibition

A world class renowned event for dialogue between research, industry, po...

COMPANY BLOGS

US Energy Grid Review Finds Needed Upgrades Would Allow More Solar,...

Yesterday (April 21) the U.S. Department of Energy released the first Qu...

Clean Energy Patents Rise in 2014, Solar Tops others, Toyota and GM...

U.S. patents for Clean Energy technologies in 2014 were again at an all ...

Environmentally friendly solutions for projects

geoAMPS joins the celebration of the worldwide observance of Earth Day o...

NEWSLETTERS

Renewable Energy: Subscribe Now

Solar Energy: Subscribe Now

Wind Energy: Subscribe Now

Geothermal Energy: Subscribe Now

Bioenergy: Subscribe Now  

 

FEATURED PARTNERS